SEPARATION OF LARGE DNA-MOLECULES WITH HIGH-VOLTAGE PULSED-FIELD GEL-ELECTROPHORESIS
SEPARATION OF LARGE DNA-MOLECULES WITH HIGH-VOLTAGE PULSED-FIELD GEL-ELECTROPHORESIS
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DOI：
10.1002/elps.11501501161
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发表时间：
1994-08-01
期刊：
影响因子：
2.9
通讯作者：
LAI, E
中科院分区：
文献类型：
--
作者：
WAGNER, L;LAI, E
我们开发了两种高压（3 kV 和 10 kV）高速脉冲场凝胶电泳系统，用于分离大至 460 kbp 的 DNA。这些系统使我们能够将高电压的快速速度与脉冲场电泳的分离能力结合起来，以实现高分辨率和短运行时间。我们发现，可以在比常用电压梯度高得多的电压梯度下分离大 DNA 片段。在 20 V/cm 下，可在 4 小时内分离大至 460 kbp 的酵母染色体，在 55 V/cm 下可在 30 分钟内快速分离 1-50 kbp DNA。对于分离相对较小的 DNA 片段，迁移率提高了 25 倍（
We have developed two high voltage (3 kV and 10 kV) high speed pulsed field gel electrophoresis systems for the separation of DNA as large as 460 kbp. These systems enable us to combine the rapid speed of high voltages and the separation power of pulsed field electrophoresis to achieve high resolution and short run durations. We found that large DNA fragments can be separated at voltage gradients much higher than commonly used. Yeast chromosomes as large as 460 kbp can be separated in 4 h at 20 V/cm and 1-50 kbp DNA can be rapidly separated in 30 min at 55 V/cm. This is 25 times faster in mobility for the separation of relatively small DNA fragments (